PAPD5 CRISPR Activation Plasmid (h): sc-412997-ACT

Human PAPD5 encodes a non-canonical poly(A) polymerase (also known as TENT4B) that resides primarily in the nucleus and participates in post-transcriptional gene regulation through RNA tailing. PAPD5 cooperates with RNA surveillance and decay machineries to influence RNA stability and quality control, shaping transcriptome homeostasis and impacting processes such as stress responses and differentiation. By modulating RNA fate, PAPD5 interfaces with broader pathways governing gene expression robustness, including ribonucleoprotein biogenesis and nuclear RNA turnover. Dysregulated PAPD5 activity has been linked in the literature to altered RNA metabolism phenotypes relevant to cancer biology and inherited disorders involving aberrant RNA processing.

PAPD5 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous PAPD5 expression without altering the underlying DNA sequence.

PAPD5 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the PAPD5 locus in human cell lines. The system is built around a catalytically inactive Cas9 (dCas9) carrying two inactivating mutations (D10A and N863A) that eliminate nuclease activity while preserving DNA binding. This dCas9 is fused to VP64, a potent transcriptional activator, and is co-expressed with a blasticidin resistance gene for selection. The second plasmid encodes the MS2-p65-HSF1 fusion protein, a secondary activator complex that works in concert with dCas9-VP64, alongside a hygromycin resistance gene. The third plasmid encodes a target-specific 20 nt sgRNA fused to two MS2 RNA aptamers that recruit the MS2-p65-HSF1 complex to the activation site, accompanied by a puromycin resistance gene. The three plasmids are delivered at a 1:1:1 mass ratio for balanced expression of all system components.

Once assembled at the target locus, the SAM complex binds within approximately 200 bp upstream of the PAPD5 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous PAPD5 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native PAPD5 locus and enabling the study of PAPD5-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of PAPD5 pathway restoration in tumor cells with silenced or reduced PAPD5 expression.

For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.